High-Content Imaging
Molecular Devices ImageXpress Micro (replaced by IXM Confocal)
For high-content automated cellular imaging and analysis we are utilizing the Molecular Devices ImageXpress Micro (datasheet - for newer model IXMicroXL) with the live cell and phase contrast/brightfield option. We can image nearly any fluorophore with out collection of Semrock filters. See below for our filter configurations.
Please contact us for information on the IXMicro. For key card access to the IXMicro room (CCSR-0116) use iLabs.
iLabs IXMicro Schedule (click here to schedule training)
View only Calendar for non-registered users
ImageXpress Micro and Catalyst Express Robotic Arm
Features and Capabilities of the ImageXpress Micro
- The IXMicro contains a fully automated inverted epifluorescence microscope -- Acquisition Guide -- User Guide
- Capable of scanning a 6-, 24-, 48-, 96- or 384-well microplates, slides or any other microplate sized container.
- Can recording multiple fluorescent images in a few minutes.
- Can quantify morphological properties through the MetaXpress analysis software (Datasheet) -- Analysis Guide.
- Our current ImageXpress Micro configuration permits the visualization of both fixed cells and live cells.
- The instrument has a temperature-, humidity-, and carbon dioxide-controlled chamber.
- Can image non-fluorescent cells/objects with the phase contrast/brightfield option
- Fully integrated to an external automated incubator (Thermo Cytomat 2C, 42 plates, Datasheet) to permit long-term or large-scale studies of cellular events via a robotic plate stacker (Thermo CataLyst Express Microplate Handler, Datasheet, User Guide, Polara Scheduling Software, Polara User Guide) thereby increasing the throughput of this workstation.
- Training provided by HTBC staff sign up here; iLabs IXMicro Schedule
- Two MetaXpress analysis computers Analysis Computer1, Analysis Computer2 also located in CCSR-0116
- Prior to training, contact Jason Wu (x5-6004) to discuss what your imaging parameters.
| DataSheet | |
| Angiogenesis tube formation | Cell health |
| Cell scoring | Count nuclei |
| Granularity | Live/dead |
| Mitotic Index | Neurite outgrowth |
| Transfluor® | Translocation — basic and enhanced |
Preferred Imaging Plates
Wells |
Vendor |
Catalog Number |
Objectives |
24 |
All |
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96 |
All |
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96 |
All |
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384 |
All |
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384 |
All |
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*Secondary Preference |
Technical Information
| Objectives | Excitation Filters | Emission Filters | Dichroics |
|---|---|---|---|
| 4X 0.2 S Fluor | 340_26nm | CUBE 1-Quad 410/504/582/669 | Quadband1 |
| 10X 0.3 Plan Fluor | 427_10nm | 440_40/521_21/607_34/700_45 | 410/504/582/669 |
| 20X 0.45 Plan Fluor ELWD | 504_12nm | CUBE 2-triple 464/542/639 | Triband |
| 40X 0.6 Plan Fluor ELWD | 589_15nm | 464_23/542_27/639_42 | 444/521/608 |
| 387_11nm | CUBE 3-CFP-LIVE | ||
| Phase Objectives | 485_20nm | 483_32 | 458 |
| 4X PhL Plan Fluor DL 0.13 NA | 560_25nm | CUBE 4-YFP-LIVE | |
| 10X Ph1 Plan Fluor DL 0.30 | 650_13nm | 542-27 | 520 |
| 20X Ph1 Plan Fluor ELWD DM | CUBE 5-HcRed-LIVE | ||
| 632-22 | Triband | ||
| FIXED Cells Profile | CUBE 6-DAPI-FIXED (Pos. 3) | ||
| 440-40 | 409 | ||
| CUBE 7-FITC-FIXED (Pos. 4) | |||
| 525-30 | 506 | ||
| CUBE 8-TRITC-FIXED (Pos. 5) | |||
| 607-36 | Quadband | ||
| FRET Profile | CUBE 9-YFP-FRET (Pos. 4) | ||
| (CFP-Live in Pos. 3) | 542-27 | 458 | |
| CUBE 10-Fura-2 (Pos. 5) | |||
| 510-84nm | Fura-2 (409) | ||
| Cy5 (Dedicated) | CUBE 11-Cy5 (Pos. 1) | ||
| 692_40nm | 660 |
Filter Sets
| Label Name | Excitation | Emission | Dichroic |
|---|---|---|---|
| Fura-2 340x | 340_26nm | D510-80nm/Fura-2 | Fura-2 |
| Fura-2 387x | 387_11nm | D510-80nm/Fura-2 | Fura-2 |
| CCF4 (FIXED) | 387_11nm | 525_30nm | 506 (FITC-FIXED) |
| Ex396/Em580 | 387_11nm | 607_36nm | Quadband (TRITC-FIXED) |
| CFP | 427_10nm | 464_23/542_27/639_42 | Triband |
| CFP-LIVE | 427_10nm | 483_32nm | 458 |
| FRET-LIVE | 427_10nm | 542_27nm | 520 (YFP-LIVE) |
| YFP-FRET | 427_10nm | 542_27nm | 458 |
| YFP | 504_12nm | 464_23/542_27/639_42 | Triband |
| YFP-LIVE | 504_12nm | 542_27nm | 520 |
| HcRED | 589_15nm | 464_23/542_27/639_42 | Triband |
| HcRED-LIVE | 589_15nm | 632_22nm | Triband |
| DAPI | 387_11nm | 440_40/521_21/607_34/700_45 | Quadband |
| DAPI-FIXED | 387_11nm | 440_40nm | 409 |
| FITC | 485_20nm | 440_40/521_21/607_34/700_45 | Quadband |
| FITC-FIXED | 485_20nm | 525_30nm | 506 |
| TRITC | 560_25nm | 440_40/521_21/607_34/700_45 | Quadband |
| TRITC-FIXED | 560_25nm | 607_36nm | Quadband |
| Ex488/Em617 | 485_20nm | 607_36nm | Quadband (TRITC-FIXED) |
| Ex510/Em580 | 485_20nm | 607_36nm | Quadband (TRITC-FIXED) |
| AlexaF594 (put HcRED_LIVE cube in Pos. 5) | 560_25nm | 632_22nm | Triband |
| FRET YFP-ALX594 (put HcRED_LIVE cube in Pos. 5) | 504_12nm | 632_22nm | Triband |
| Cy5 | 650_13nm | 440_40/521_21/607_34/700_45 | Quadband |
| Cy5 | 650_13nm | 692_40nm | 660 |
Optimized for these fluorophores
Cyan: CFP, AmCyan, SYTOX Blue, BOBO-1, BO-PRO-1
Yellow: YFP, Calcium Green-1, Fluo-3, Rhodamine 123
Red: HcRed, Cy3.5, Texas Red®, Alexa Fluor 594
Set description
This 5-filter "Pinkel" set is perfectly optimized for imaging CFP, YFP, and HcRed with high brightness, extremely low crosstalk, and superb signal-to-noise ratio. The set is also ideal for samples triple-labeled with CFP, YFP, and an orange or red dye, like Texas Red. It is comprised of three single-band exciters along with a triple-band beamsplitter and a triple-band emitter, and thus works best with an exciter filter wheel or slider.
Actual measured data from typical filters
| Filter | Average Transmission | Center Wavelengths | Bandwidths * |
| Exciter 1 | > 90% | 427 nm | 10 nm |
| Exciter 2 | > 90% | 504 nm | 12 nm |
| Exciter 3 | > 90% | 589 nm | 15 nm |
| Emitter | > 90%, 90%, 90% |
464, 542, 639 nm | 23, 27, 42 nm |
| Average Reflection | Reflection Bands | Transmission Bands | |
| Dichroic | > 95% > 95% > 95% |
420 - 430 nm 496 - 510 nm 579 - 596 nm |
451 - 480 nm 530 - 561 nm 618 - 664 nm |
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Exciter 1
Dichroic
Emitter
Exciter 2
Exciter 3
Optimized for these fluorophores
Blue: DAPI, Hoechst, AMCA, Alexa Fluor 350
Green: FITC, GFP, rsGFP, BODIPY, Alexa Fluor 488
Orange: TRITC, Cy3™, Texas Red®, MitoTracker Red,
Alexa Fluor 568 & 594
Red: Cy5™, APC, TOTO-3, TO-PRO-3, Alexa Fluor 647 & 660
Set description
This 6-filter "Pinkel" quad-band set is designed for sequentially imaging DAPI, FITC, TRITC, and Cy5 (and other key fluorophores – see complete list above). This state-of-the-art set represents a break-through in quad-band performance, and has been tested by independent researchers to have up to four times the brightness, twice the signal-to-noise ratio and substantially lower crosstalk than any other such quad-band filter set on the market. The complete set is comprised of a quad-band beamsplitter and a quad-band emitter (these can be offered assembled in a microscope cube), plus four “no burn-out” single-band exciters that are intended to be mounted in an exciter filter wheel or slider.
Actual measured data from typical filters
| Filter | Average Transmission | Center Wavelengths | Bandwidths * |
| Exciter 1 | > 85% | 387 nm | 11 nm |
| Exciter 2 | > 90% | 485 nm | 20 nm |
| Exciter 3 | > 90% | 560 nm | 25 nm |
| Exciter 4 | > 90% | 650 nm | 13 nm |
| Emitter | > 90%, 90%, |
440, 521, 607, 700 nm |
40, 21, 34, 45 nm |
| Average Reflection | Reflection Bands | Transmission Bands | |
| Dichroic | > 95% > 95% > 95% > 95% |
381 - 392 nm 475 - 495 nm 547 - 572 nm 643 - 656 nm |
420 - 460 nm 510 - 531 nm 589 - 623 nm 677 - 722 nm |
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Exciter 4